Human RNF2 (RING2 / RING1B) knockout HeLa cell line
- Advanced Validation
- What is this?
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RNF2 KO cell line available to order. KO validated by. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 8 bp deletion in exon 2 and Insertion of the selection cassette in exon 2.
View Alternative Names
DING, DinG protein, E3 ubiquitin protein-ligase RING 2, E3 ubiquitin-protein ligase RING2, HIP2-interacting protein 3, HIPI 3, Huntingtin-interacting protein 2-interacting protein 3, OTTHUMP00000060668, Polycomb M33 interacting protein Ring 1B, Protein DinG, RING 1B, RING finger protein 1B, RING finger protein 2, RING finger protein BAP-1, RING2_HUMAN, RNF 2
- WB
Lab
Western blot - Human RNF2 (RING2 / RING1B) knockout HeLa cell line (AB264845)
Lanes 1- 2 : Merged signal (red and green). Green - ab181140 observed at 42 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) observed at 50 kDa.
ab181140 was shown to react with RING2 / RING1B / RNF2 in wild-type HeLa cells in western blot. The band observed in knockout cell line ab264845 (knockout cell lysate ab257640) lane below 42kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type HeLa and RNF2 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab181140 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-RING2 / RING1B / RNF2 antibody [EPR12245] (<a href='/en-us/products/primary-antibodies/ring2-ring1b-rnf2-antibody-epr12245-ab181140'>ab181140</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
RNF2 CRISPR/Cas9 edited HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human RNF2 (RING2 / RING1B) knockout HeLa cell line (ab264845)
Predicted band size: 37 kDa
Observed band size: 42 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human RNF2 (RING2 / RING1B) knockout HeLa cell line (AB264845)
Allele-1 : 8 bp deletion in exon 2.
- Sanger seq
Unknown
Sanger Sequencing - Human RNF2 (RING2 / RING1B) knockout HeLa cell line (AB264845)
Allele-2 : Insertion of the selection cassette in exon 2.
Reactivity data
Product details
Recommended control: Human wild-type HeLa cell line (ab255448). Please note a wild-type cell line is not automatically included with a knockout cell line order, if required please add recommended wild-type cell line at no additional cost using the code WILDTYPE-TMTK1.
We will provide viable cells that proliferate on revival.
Western blot data indicates that the CRISPR gene edit may have resulted in a truncation of the protein of interest. Please see data images.
This product is subject to limited use licenses from The Broad Institute, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and patents please refer to our limited use license and patent pages.
What's included?
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
Handling procedures
Initial handling guidelines
Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80°C. Storage at -80°C may result in loss of viability.
1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.
2. Transfer the cell suspension (0.8 mL) to a 15 mL/50 mL conical sterile polypropylene centrifuge tube containing 8.4 mL pre-warmed culture medium, wash vial with an additional 0.8 mL culture medium (total volume 10 mL) to collect remaining cells, and centrifuge at 201 x g (rcf) for 5 minutes at room temperature. 10 mL represents minimum recommended dilution. 20 mL represents maximum recommended dilution.
3. Resuspend the cell pellet in 5 mL pre-warmed culture medium and count using a haemocytometer or alternative cell counting method seed all remaining cells into a T25.
4. Incubate the culture at 37°C incubator with 5% CO2. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.
5. Once confluent passage into an appropriate flask at a density of 2x104 cells/cm2. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.
Subculture guidelines
- All seeding densities should be based on cell counts gained by established methods.
- A guide seeding density of 2x104 cells/cm2 is recommended.
- Cells should be passaged when they have achieved 80-90% confluence.
Culture medium
DMEM (High Glucose) + 10% FBS
Cryopreservation medium
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
RING2/RNF2/RING1B regulates gene expression by modifying chromatin structure especially through monoubiquitination of histone H2A at lysine 119. This action occurs within the Polycomb Repressive Complex 1 a multiprotein structure important for maintaining the transcriptional repression of target genes. It influences cell cycle progression and development by silencing genes involved in proliferation and differentiation.
Pathways
RING2/RNF2/RING1B's role connects to the Polycomb group (PcG) pathway and the Wnt signaling pathway. Within these pathways it interacts with proteins like BMI1 and EZH2 which are other components of polycomb complexes. These interactions help modulate chromatin dynamics and control the expression of several genes involved in cell identity and development.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Target data
Complementary Products
Primary Antibodies
AB181140
Anti-RING2 / RING1B / RNF2 antibody [EPR12245]
RabMAb|Recombinant|KO Validated
![Western blot - Anti-RING2 / RING1B / RNF2 antibody [EPR12245] (AB181140)](https://content.abcam.com/products/images/ring2-ring1b-rnf2-antibody-epr12245-ab181140--western-blot-img148168.jpg)
- 4
- 3
Primary Antibodies
AB7753
Anti-pan Cytokeratin antibody [C-11]
BOND RX™ Validated|Recombinant|Lab Essentials|Advanced Validation
![Immunohistochemistry (Frozen sections) - Anti-pan Cytokeratin antibody [C-11] (AB7753)](https://content.abcam.com/products/images/pan-cytokeratin-antibody-c-11-ab7753--immunohistochemistry-frozen-sections-img31615.jpg)
- 4
- 23
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
For licensing inquiries, please contact partnerships@abcam.com